Malaria is a leading cause of human death and illness, causing over 300 million cases of clinical malaria and 1+ million deaths each year. Traditional measures to control and cure malaria are becoming increasingly less effective and there is an urgent need for the development of new drugs and vaccines. A strategic hurdle for development of new anti-malarial therapeutics remains the lack of experimentally validated functional information about most P. falciparum genes. Critical for identification of new drug targets is a better understanding of essential metabolic pathways and weaknesses in the parasite's physiology. Our approach will create a large library of P. falciparum parasite mutant clones with single piggyBac insertions that will be used for phenotypic analysis. Preliminary studies have validated this approach to be able to identify processes critical for blood-stage growth. Through this project we expect to identify dozens of genes critical for parasite virulence and development processes that are novel targets on which drug discovery projects can be initiated. An additional important outcome of our project will provide the malaria research community with a large collection of gene knockouts that will be a valuable resource for a multitude of other research projects. PUBLIC HEALTH RELEVANCE: Malaria is a devastating global health problem and its elimination as an important disease requires new therapies. A major hurdle for development of new an anti-malarial drug is a lack of comprehensive information about the best targets. New drugs effective against multiple different targets in malaria parasites are essential to effectively eliminate malaria. Our project will provide this type of knowledge to identify new drug targets.